Experimental Pharmacology-I 29

Experiments
EXPERIMENT NO. 3
C OF DRUGS ON BLOOD PRESSURE AND HEART RATE OF
DOG
Purpose:
At the end of practical class, the students shallbe able to
1. Know about blood pressure and its significance.
Know the effects of different drugs on the blood pressure and heart rate.
3. Know about measuring of blood pressure of dog by using mercury manometer.
Terminology:
Blood pressure: The arterial blood pressure is defined as the pressure exerted by the
blood on the walls of the blood vessels. Blood pressure is usually expressed in terms
of the systolic pressure (maximum during one heart beat) or diastolic pressure
(minimumin between two heart beats) and is measured in millimeters of mercury
(mmHg), above the surrounding atnospheric pressure. Arterial blood pressure is
measured with a sphygmomanometer and is usually 120/80 mmHg or 16/11 kPa
(kilopascals). Blood pressure = Cardiac output x Peripheral resistance.
Systolic pressure: When the left ventricle contracts and pushes blood into the aota
the pressure produced within the arterial system is called the systolic blood pressure.
In adults it is about 120 mmHg or 16 KPa.
Diastolic pressure: When complete cardiac diastole occurs and the heart is resting
following the ejection of blood, the pressure within the arteries is called diastolic
blood pressure. In an adult, this is about 80 mmHg or 11 kPa.
Pulse pressure: The difference between systolic and diastolic blood pressures is the
pulse pressure. It is approximately about 40 mmHg.
Peripheral resistances: Vascular resistance to the flow of blood in peripheral arterial
vessels is known as peripheral resistances. It depends on the factors like internal
vesseldiameter, vessel length and blood viscosity.
Cardiac output: The amount of blood ejected out of the heart in one minute is
known as cardiac output. The cardiac output is determined by the stroke volume and
the heart rate.
Stroke volume: The amount of blood ejected out of each ventricle in one beat is
known as stroke volume.
Description:
Tne systemic arterial blood pressure, usually called arterial blood pressure, is the result of
The discharge of blood from the left ventricle into the already fullaorta. These figures varyat
according to the time of day, the posture, gender and age of anindividual. During bedrest
night the blood pressure tends to be lower. It increases with age and is usually higher in
Women than in men.
Experimental Pharmacology-I 30

animal like
Experime
dog, cat or rabbit is EX
standardof the
of theRecording methods pressure in an anaesthetized
bloodfor the pharmacological studies of drugs. Blood pressure is studiob
mercury
1828 originally designed the obtained
by Poiseuille's manometer. Poissuille in
be
manometer an Pri
the merCury,
later in 1847 so Ludwig
Carlthat modified it to allow graphic
records to from afloat d
progressive changes in the blood pressure could be studied Now aldkh
manometer with
basically same. It consists of a mercury
the assembly remains the smoked surface ofthe kymograph
withthe
Supporting a long stiff wire that write on
the mean arterial helu
pressure. The mar
assembly only records
of a writing point. The glass tube with two vertiCal limos about 30 cm in hein
manometer consists of a U- shaped
filled with mercury. The mercury is displaced equally und
and 5 mm bore. Half of the tube is obtain the actual pressure in terms of a mare
other, hence to
one limb and down in the multiplied by two. To avoid this, a milimetd
recorded must be
column, the displacement fitted with the manometer. On the Surface def
mmHg is
scale with doubled values, up to 250 float supporting a long stiff wire, bearing ar
column has a cylindrical
one limb of the mercury rotating smoked paper surtace of the kymograot R
writes on the
stylus on its upper end that side-tube that is connected through
inextensibi
of the manometer has a
The other limb arterial cannula. The upper end of the limd
rubber or polythene to an
tube made up of thick that can bie
connected with a reservoir bottle containing some anticoagulant fluid
is also interconnecting tube to the arterial cannula. At the
pumped into this limb and through the
connects or disconnects the arterial cannu p
junction of the side tube, a three way stopcock
3.1).
and manometer from the reservoir bottle (Fig.

the effect of drugs on the blood pressure of dog using mercury

Fig. 3.1:Study
manomneter
 Experimental Pharmacology-I 31 Experiments

Objective: Toostudy the effect of various drugs on blood pressure and heart rate of dog.
Principle:
Blood pressure is the torce or pressure which the circulating blood exerts on the walls of
the blood vessels. It may be systolic or diastolic pressure. Blood pressure is directly
proportional to cardiac output and peripheral resistance. The heart and blood vessels are
under the control of autonomic nervous system. Sympathetic stimulation increases the
cardiac output and peripheral' resistances and increases the blood pressure. In contrast,
parasympathetic stimulation decreases the cardiac output, which lowers the blood pressure.
Drugs which increase the blood pressure are called pressor agents and those decrease the
'bloodpressure are called depressor agents.
There are several drugs which influence the blood pressure and heart rate, and their
effects can be studied by using anaesthetized dog. Blood pressure in an anaesthetized
animal can be recorded by using Poiseuille's mercury manometer as described above.
Requirements:
Anima/tissue: Dog / Dog heart.
Drugs: Heparin, saline (0.9% NaC), pentobarbitone sodium, adrenaline, noradrenaline,
isoprenaline, propranolol, acetylcholine, atropine, histamine.
Equipment: Kymograph, arterial cannula, venous cannula, tracheal cannula, 'U' shaped or
Poiseuille's mercury monometer.
Others: Stand, syringe, beaker, stop watch, dissection box, thread, and connecting rubber
tubing.
Procedure:
1. Prepare the stock solution of different drugs to study the effect on blood pressure
and heart rate on anaesthetized dog.
2. Weigh an adult healthy dog and anaesthetize it with pentobarbitone sodium (25
mg/kg, intravenously).
3. Fix the animal onthe operation table by tying its legs. Expose the trachea and two
carotid arteries by making a midline incision in the neck region.
4. Insert a tracheal cannula into the trachea to ensure a free airway and to provide
artificial respiration if necessary. Effect of drugs on respiration can also be recorded.
5. Cannulate one carotid artery by an arterial cannula through asmall incision and
Connected via a three way stopcock to the U Poiseuille's manometer and to the
pressure bottle of the mercury manometer system filled with 0.9% NaCl solution
containing heparin (anticoagulant, 1000 units per m). The space between manometer
and the arterial cannula is also filled with saline-heparin solution and care should be
taken to avoid entry of any air bubble.
b. Pass a thread under another carotid artery and clamp it to study the effect of carotid
occlusion.
Experimental Pharmacology-II 32
Experimen
7. Dissect the femoral vein from femoral triangle and cannulate it with a thin polythe
tube connected via athree way stopcock for injecting various drugs and for flushin
with saline-heparin solution after each injection.
cannula and record their effect o
8. Inject different drugs one by one through venous
the blood pressure of dog.
Observation:
pressure and heart rate of dog
Table 3.1: Effect of various drugs on blood
Effects on
Drugs (Concentration of stock Dose (ug) Blood pressure Heart rate
solution) Beatslmin)
NO (mmHg)

1. Normal
2
2. Adrenaline (10 ug/ml)
2
3. Noradrenaline (10 ug/ml)
2
4. Isoprenaline (10 ug/ml)
Propranolol (1 mg/m) 200
5.
2
6. Acetylcholine (10 ug/ml)
20
7. Atropine(100 ug /ml)
2
Histamine (10 ug/ml)
0.2 ml
Volume of above solutions to be injected =
Inference:
vasoconstriction and increased systoli
Noradrenaline (predominantly a agonist) causes
diastolic pressure, with areflex bradycardia. Only rise in blood pressure is seen and th
and
vasoconstriction of blood vessels specially those which supply blood to skin and
is due to the rise in B.P. is more tha
not seen, the effect of
mucosa. Since B action of noradrenaline is
adrenaline.
isoprenaline, aselective Bagonist, is a vasodilator, but strongly increase
Isoproterenolor result o
and rate. B1 receptor present in heart are excited by isoprenaline as a
cardiac force
in the heart rate and blood pressure. When drug reaches periphen
which there is increase muscle by
vasodilation of blood vessels specially which are supplied to skeletal
there occurs
receptors. As aresult mean arterial pressure falls.
acting onB2 through a and B receptors. As
actions i.e. produces effect
Adrenaline combines both rate and force O
stimulation there is sudden increase in heart
result of B1 receptor due to reflex inhibition, there is slight decrease but drug reache
contraction. Immediately stimulation produces vasoconstriction
and hence. furthe
where , receptor
periphery of adrenaline is reduce
the blood pressure is observed. When the concentration
rise in
Experimental Pharmacology-I 33 Experiments

Ro receptor action predominates and as a result slight fall in blood pressure (secondary tall) Is
Seen.
cardiac
Badrenergic receptor blockers like propranolol causes little change in heart rate,
to
output or arterial pressure. In presence of propranolol the sympathomimetic drugs fails
produce any effects on blood pressure.
Acetylcholine is parasympathomimetic agent which produces its action on heart
cardiac
through muscarinic (M2) receptors. These include cardiac slowing and a decrease in
of the atria,
output. The latter action is mainly due to a decreased force of contraction
because the ventricles have only asparse parasympathetic innervation and a low sensitivity
occurs (a nitric oxide-mediated
to muscarinic agonists. Generalised vasodilatation also
effect),and the above two effects combine to produce asharp fall in arterial pressure.
cardiac
Atropine causes tachycardia through blocking muscarnic (M) receptors of
only inhibition of
muscle. This is because there is no effect on the sympathetic system, but
atropine causes a paradoxical
the existing parasympathetic tone. At very low doses, unaffected, because
bradycardia, possibly due to acentral action. Arterial blood pressure is
of atropine, ACh fails to
most resistance vessels have no cholinergic innervation. In presence
produce any action.
generalised vasodilation
Histamine is a naturally occurring autacoid which causes
the dog there is marked fall
through HË and H, receptors. When histamine is administered in
in blood pressure.
Precautions:
1. Handle the dog with care.
2. Maintain the aseptic condition of the laboratory.
stress.
3. Acclimatize the animals to the laboratory prior to testing for minimizing the
4. Interpretation should be carried out by experienced
person.
Questions for the students:
1. Define blood pressure.
2. Discuss the experimentscarried out by using dog.
3. List out the drugs which increase or decrease the blood pressure.
4. Discuss the effects of sympathomimetic and parasympathomimetic drugs on the
blood pressure.